摘要
目的探索胰蛋白酶分离香猪角质形成细胞膜片的最佳方案,在胶原-壳聚糖人工真皮支架创面上进行角质形成细胞体内培养,明确该方法修复猪创伤全层皮肤缺损的实用性。方法用不同皮肤厚度、胰蛋白酶浓度和分离温度分离出角质形成细胞,把有活力的角质形成细胞接种于人工真皮支架植入后1周的全层皮肤缺损创面,2周后对创面愈合进行观察并作病理检测。结果在37℃条件下用0.20%的胰蛋白酶消化1mm厚度皮片100分钟为最佳分离方案。2周后肉眼和病理切片均显示接种的角质形成细胞膜片成活并完全覆盖修复创面。结论应用角质形成细胞体内培养加人工真皮支架制成"人工皮肤"能较好的修复猪创伤全层皮肤缺损,抑制创面收缩和瘢痕形成。
Objective To explore the best method for separating epidermis films from dermis in Bama miniature pigs with trypsin and to culture keratinocyte in vivo on surfaces of artificial dermal stent which had been transplanted on wounds of full-thickness skin defect,and to determine the practicality of full-thickness skin defect repair.Methods Epidermis films were tried to separate from dermis with trypsin by regulating skin thickness of harvested skin,digestion temperature and concentrations of trypsin.The separated epidermis films after vigor detection were then transplanted on surfaces of artificial dermal stent which had been transplanted on wounds of full-thickness skin defect for 1 week and cultured in vivo.Wound surfaces were observed and samples were harvested in 2 weeks after epidermis films were transplanted on surfaces of artificial dermal stent.All harvested samples were pathologically observed.Results It was the most suitable method for separating keratinocytes from dermis to digest 1 mm thickness skin with 0.2% trypsin for 100 minutes under 37℃.Keratinocytes which had been grafted on surface of artificial dermal stent for 2 weeks could perfectly survive,and the keratinocytes gradually covered the wounds until all surfaces of wounds were completely covered by keratinocytes.Epidermis perfectly survived through pathological observation.The artificial skin＇s structure is similar to normal skin.Conclusion Keratinocytes culture in vivo combined with artificial dermal stent application played a very important role in wound repair through preventing wound contraction and scar formation.
出处
《浙江创伤外科》
2011年第2期151-154,共4页
Zhejiang Journal of Traumatic Surgery
基金
浙江省医药卫生科学研究基金A类(2008A131)
